Many of the targets of drug discovery efforts are proteins, including membrane proteins. To date, the ability to accumulate relevant structural and functional information for membrane proteins through the generation of ligand-protein complex structures has been limited. And the inability to generate these structures is in part due to the limitations that arise because of the requirement for the use of detergents to extract, purify and crystallize membrane proteins.
To this end, there has been recent progress in generating crystal structures of seven-transmembrane, G-protein coupled receptors (GPCR's) such as β1 and β2-adrenergic receptors (Warne, et al., 2008; Hanson, et al., 2008)) the adenosine A(2A) receptor (Jaakola, et al., 2008) and the glucagon receptor, GCGR (siu, et al., 2013). However, these structures were generated using highly mutated forms of the proteins, including multiple point mutations and the insertion of a large fusion domains; and these mutations often alter the ligand binding characteristics of these proteins. Additionally, the crystals used to generate these GPCR structures were grown using the technique of lipidic-cubic phase crystallization which is limited in its utility due to the difficulties in handling the lipid-protein mixture, visualizing crystals within the lipid matrix and retrieving the small crystals from the lipid matrix (Landau & Rosenbusch 1996; Nollert 2004).
Clearly, there is a need for additional techniques or methods to gain increased structural understanding of wild-type membrane proteins. Ideally, one would like to identify a robust crystallization technique that would allow for the crystallization of fully functional, wild-type membrane proteins without the presence of detergents, e.g., using vapor-diffusion crystallization.